The present invention relates to lipolytic enzymes, methods of using and producing lipolytic enzymes, as well as nucleic acid sequence encoding same.
Lipolytic enzymes (such as lipases and phospholipases) are capable of hydrolyzing carboxylic ester bonds in a substrate to release carboxylic acids. They are known to be useful, e.g., in baking and detergents.
A lipase from Fusarium heterosporum and its sequence are known. Journal of Fermentation and Bioengineering, 75 (5), p 349-352, 1993. Journal of Biochemistry (Tokyo), 116 (3), p 536-540, 1994.
Some isolates of Acremonium are known to produce lipase. Roberts et al., Mycologia, 79, 265-273, 1987. Satyanarayana and Johri, Current Science, 50, 680-682, 1981.
The inventors have isolated a lipolytic enzyme from a strain of Acremonium berkeleyanum. The inventors also isolated the gene encoding the novel lipolytic enzyme and cloned it into an E. coli strain.
Accordingly, the invention provides a lipolytic enzyme which may be a polypeptide having an amino acid sequence as the mature peptide shown in SEQ ID NO: 1.
Further, the lipolytic enzyme of the invention may be a polypeptide encoded by the lipolytic enzyme encoding part of the DNA sequence cloned into a plasmid present in Escherichia coli deposit number DSM 13538.
The lipolytic enzyme may also be an analogue of the polypeptide defined above which:
i) has at least 70% homology with said polypeptide,
ii) is immunologically reactive with an antibody raised against said polypeptide in purified form,
iii) is an allelic variant of said polypeptide,
The lipolytic enzyme of the invention may also be a polypeptide which is encoded by a nucleic acid sequence which hybridizes under high stringency conditions with a complementary strand of the nucleic acid sequence of SEQ ID NO: 1 encoding the mature polypeptide or a subsequence thereof having at least 100 nucleotides.
The nucleic acid sequence of the invention may comprise a nucleic acid sequence which encodes the lipolytic enzyme described above. The nucleic acid sequence may also comprise:
a) the DNA sequence encoding a mature lipolytic enzyme cloned into a plasmid present in Escherichia coli DSM 13538,
b) the DNA sequence encoding a mature lipolytic enzyme shown in SEQ ID NO: 1, or
c) an analogue of the DNA sequence defined in a) or b) which
i) has at least 70% homology with said DNA sequence, or
ii) hybridizes at high stringency with said DNA sequence, its complementary strand or a subsequence thereof.
Other aspects of the invention provide a recombinant expression vector comprising the DNA sequence, and a cell transformed with the DNA sequence or the recombinant expression vector.
A comparison with full-length prior-art sequences shows that the mature amino acid sequence of the invention has 67% homology with the lipase from Fusarium heterosporum described above, and the corresponding DNA sequence of the invention shows 68% homology with that of the F. heterosporum enzyme.